Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat (predicted:Pig,Cow,Horse,Rabbit)

IgG

Human,Mouse,Rat (predicted:Pig,Cow,Horse,Rabbit)

1. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-phospho-AMPK alpha 2 (Ser173) antibody (TMAB-01389)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C, The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
2. Sample:
Muscle (Mouse) Lysate at 40 μg
Heart (Mouse) Lysate at 40 μg
Primary: Anti-phospho-AMPK alpha 2 (Ser173) (TMAB-01389) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
3. Paraformaldehyde-fixed, paraffin embedded (rat heart tissue); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (p-AMPK alpha 2) Polyclonal Antibody, Unconjugated (TMAB-01389) at 1:400 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
4. Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-phospho-AMPK alpha 2 (Ser173) antibody (TMAB-01389)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2ug/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human AMPK alpha 2 around the phosphorylation site of Ser173

Human

5563

Integration of energy metabolism,Response to hypoxia,Metabolism,Integration of energy,Fatty acids,Hypoxia,Fatty acid oxidation,Metabolism,Other Kinases